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Tirtha Datta

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Papers by Tirtha Datta

Identification of protein candidates in spermatozoa of water buffalo (Bubalus bubalis) bulls helps in predicting their fertility status

Frontiers in Cell and Developmental Biology

The water buffalo (Bubalus bubalis) is an indispensable part of the Indian dairy sector and in se... more The water buffalo (Bubalus bubalis) is an indispensable part of the Indian dairy sector and in several instances, the farmers incur economic losses due to failed pregnancy after artificial insemination (AI). One of the key factors for the failure of conception is the use of semen from the bulls of low fertilizing potential and hence, it becomes important to predict the fertility status before performing AI. In this study, the global proteomic profile of high fertile (HF) and low fertile (LF) buffalo bull spermatozoa was established using a high-throughput LC-MS/MS technique. A total of 1,385 proteins (≥1 high-quality PSM/s, ≥1 unique peptides, p < 0.05, FDR < 0.01) were identified out of which, 1,002 were common between both the HF and LF groups while 288 and 95 proteins were unique to HF and LF groups respectively. We observed 211 and 342 proteins were significantly high (log Fc ≥ 2) and low abundant (log Fc ≤ 0.5) in HF spermatozoa (p < 0.05). Gene ontology analysis revea...

Establishment of a repertoire of fertility associated sperm proteins and their differential abundance in buffalo bulls (Bubalus bubalis) with contrasting fertility

Scientific Reports

Sperm harbours a wide range of proteins regulating their functions and fertility. In the present ... more Sperm harbours a wide range of proteins regulating their functions and fertility. In the present study, we made an effort to characterize and quantify the proteome of buffalo bull spermatozoa, and to identify fertility associated sperm proteins through comparative proteomics. Using high-throughput mass spectrometry platform, we identified 1305 proteins from buffalo spermatozoa and found that these proteins were mostly enriched in glycolytic process, mitochondrial respiratory chain, tricarboxylic acid cycle, protein folding, spermatogenesis, sperm motility and sperm binding to zona pellucida (p < 7.74E−08) besides metabolic (p = 4.42E−31) and reactive oxygen species (p = 1.81E−30) pathways. Differential proteomic analysis revealed that 844 proteins were commonly expressed in spermatozoa from both the groups while 77 and 52 proteins were exclusively expressed in high- and low-fertile bulls, respectively. In low-fertile bulls, 75 proteins were significantly (p < 0.05) upregulated...

Analysis of amplification and association polymorphisms in the bovine beta-defensin 129 (BBD129) gene revealed its function in bull fertility

Scientific Reports

β-defensins are adsorbable on the sperm surface in the male reproductive tract (MRT) and enhance ... more β-defensins are adsorbable on the sperm surface in the male reproductive tract (MRT) and enhance sperm functional characteristics. The beta-defensin 129 (DEFB129) antimicrobial peptide is involved in sperm maturation, motility, and fertilization. However, its role in bovine fertility has not been well investigated. This study examines the relationship between the bovine BBD129 gene and Bos indicus x Bos taurus bull fertility. The complete coding sequence of BBD129 mRNA was identified by RNA Ligase Mediated-Rapid Amplification of cDNA End (RLM-RACE) and Sanger sequencing methodologies. It consisted of 582 nucleotides (nts) including 5' untranslated region (UTR) (46nts) and 3'UTR (23nts). It conserves all beta-defensin-like features. The expression level of BBD129 was checked by RT-qPCR and maximal expression was detected in the corpus—epididymis region compared to other parts of MRT. Polymorphism in BBD129 was also confirmed by Sanger sequencing of 254 clones from 5 high fert...

Shotgun Proteomics-based distinctive fingerprint of buffalo sperm surface unravels over-representation of proteins driving spermatogenesis, motility, and fertility

The mammalian sperm surface is radically modified in its biomolecular composition and structure d... more The mammalian sperm surface is radically modified in its biomolecular composition and structure during the transit of sperm through the male and female reproductive tract (MRT, FRT). Numerous distinct secretagogues such as (glyco)proteins and many GPI-anchored proteins (GPI-APs) are applied as peripheral coats on the sperm surface in the MRT, which is critical for the maturation, survival and function of the spermatozoa. This study aimed to characterize the buffalo sperm surface proteins by extracting them either by using elevated salt (NaCl) or Phosphatidylinositol-specific phospholipase-C (for GPI-APs) or by inducing in vitro capacitation. A buffalo sperm surface-specific proteomic fingerprint was subsequently generated using the protein informatics platform, Proteome Discoverer (v2.2) for protein identification. Overall, 1695 unique proteins isoforms (proteoforms) (≥ 1 high-quality PSM/s, minimum 2 peptides, P < 0.05, FDR < 0.01) that exhibited remarkable heterogeneity in t...

Cryopreservation process alters the expression of genes involved in pathways associated with the fertility of bull spermatozoa

Frontiers in Genetics

In bovines, cryopreserved semen is used for artificial insemination; however, the fertility of cr... more In bovines, cryopreserved semen is used for artificial insemination; however, the fertility of cryopreserved semen is far lower than that of fresh semen. Although cryopreservation alters sperm phenotypic characteristics, its effect on sperm molecular health is not thoroughly understood. The present study applied next-generation sequencing to investigate the effect of cryopreservation on the sperm transcriptomic composition of bull spermatozoa. While freshly ejaculated bull spermatozoa showed 14,280 transcripts, cryopreserved spermatozoa showed only 12,375 transcripts. Comparative analysis revealed that 241 genes were upregulated, 662 genes were downregulated, and 215 genes showed neutral expression in cryopreserved spermatozoa compared to fresh spermatozoa. Gene ontology analysis indicated that the dysregulated transcripts were involved in nucleic acid binding, transcription-specific activity, and protein kinase binding involving protein autophosphorylation, ventricular septum morph...

Sperm Transcripts Associated With Odorant Binding and Olfactory Transduction Pathways Are Altered in Breeding Bulls Producing Poor-Quality Semen

Frontiers in Veterinary Science

Spermatozoa carries a reservoir of mRNAs regulating sperm functions and fertilizing potential. Al... more Spermatozoa carries a reservoir of mRNAs regulating sperm functions and fertilizing potential. Although it is well recognized that a considerable proportion of high genetic merit breeding bulls produce poor-quality semen, the transcriptomic alterations in spermatozoa from such bulls are not understood. In the present study, comparative high-throughput transcriptomic profiling of spermatozoa from good and poor-quality semen-producing bulls was carried out to identify the transcripts associated with semen quality. Using next-generation sequencing (NGS), we identified 11,632 transcripts in Holstein Friesian bull spermatozoa; after total hit normalization, a total of 544 transcripts were detected, of which 185 transcripts were common to both good and poor-quality semen, while 181 sperm transcripts were unique to good quality semen, and 178 transcripts were unique to poor-quality semen. Among the co-expressed transcripts, 31 were upregulated, while 108 were downregulated, and 46 were neu...

Acute Exposure to Organophosphosphorus Pesticide Metabolites compromises mammalian sperm survival, functional and fertility

Agrichemicals such as metabolites (OPPMs) of the commonly-used organophosphorus pesticides (OPPs)... more Agrichemicals such as metabolites (OPPMs) of the commonly-used organophosphorus pesticides (OPPs) are considered more hazardous and pervasive than their parent pesticides which. This is because parental germline exposure to such xenobiotics has been demonstrated to cause an elevated susceptibility towards various reproductive failures e.g. sub- or in-fertility Buffalo is a vital livestock resource, however, has numerous reproductive restraints e.g. male subfertility (low conception) that negatively impact its lifetime productivity. This study sought to examine the effects of low-dose, acute OPPM exposure on the mammalian sperm survival and its fertilizing ability. The buffalo spermatozoa were briefly exposed to the three most prevalent organophosphorus pesticides metabolites viz. Omethoate (Dimethoate), paraoxon-methyl (methyl parathion) and 3, 5, 6-trichloro-2-pyridinol, (chlorpyrifos). The structural and functional integrity of the OPPM-exposed and control sperm were evaluated by ...

Integrated multi-omics analyses reveals molecules governing sperm metabolism potentially influence bull fertility

Scientific Reports

Bull fertility is of paramount importance in bovine industry because semen from a single bull is ... more Bull fertility is of paramount importance in bovine industry because semen from a single bull is used to breed several thousands of cows; however, so far, no reliable test is available for bull fertility prediction. In the present study, spermatozoa from high- and low-fertility bulls were subjected to high-throughput transcriptomic, proteomic and metabolomic analysis. Using an integrated multi-omics approach the molecular differences between high- and low-fertility bulls were identified. We identified a total of 18,068 transcripts, 5041 proteins and 3704 metabolites in bull spermatozoa, of which the expression of 4766 transcripts, 785 proteins and 33 metabolites were dysregulated between high- and low-fertility bulls. At transcript level, several genes involved in oxidative phosphorylation pathway were found to be downregulated, while at protein level genes involved in metabolic pathways were significantly downregulated in low-fertility bulls. We found that metabolites involved in T...

Comparative Proteome Profiling of Saliva Between Estrus and Non-Estrus Stages by Employing Label-Free Quantitation (LFQ) and Tandem Mass Tag (TMT)-LC-MS/MS Analysis: An Approach for Estrus Biomarker Identification in Bubalus bubalis

Frontiers in Genetics

Accurate determination of estrus is essentially required for efficient reproduction management of... more Accurate determination of estrus is essentially required for efficient reproduction management of farm animals. Buffalo is a shy breeder and does not manifest overt signs of estrus that make estrus detection difficult resulting in a poor conception rate. Therefore, identifying estrus biomarkers in easily accessible biofluid such as saliva is of utmost interest. In the current study, we generated saliva proteome profiles during proestrus (PE), estrus (E), metestrus (ME), and diestrus (DE) stages of the buffalo estrous cycle using both label-free quantitation (LFQ) and labeled (TMT) quantitation and mass spectrometry analysis. A total of 520 proteins were identified as DEPs in LFQ; among these, 59 and four proteins were upregulated (FC ≥ 1.5) and downregulated (FC ≤ 0.5) during E vs. PE, ME, and DE comparisons, respectively. Similarly, TMT-LC-MS/MS analysis identified 369 DEPs; among these, 74 and 73 proteins were upregulated and downregulated during E vs. PE, ME, and DE stages, respe...

Additional file 3 of Buffalo sperm surface proteome profiling reveals an intricate relationship between innate immunity and reproduction

Additional file 3: Supplementary Figure 3. The removal of glycans after salt and PI-PLC treatment... more Additional file 3: Supplementary Figure 3. The removal of glycans after salt and PI-PLC treatment. Overlay of the MFI histograms obtained by flow cytometry analysis of control, elevated salt and PI-PLC treated spermatozoa from buffalo bulls (n = 3 incubated with six FITC-labelled lectins viz. ABL (A) JAC (B) LEL (C), LCA(D), MAL-II(E) AND PNA(F).

Additional file 7 of Buffalo sperm surface proteome profiling reveals an intricate relationship between innate immunity and reproduction

Additional file 7: Supplementary sheet-Results. Identified sperm-surface proteins and their gene ... more

Additional file 1 of Buffalo sperm surface proteome profiling reveals an intricate relationship between innate immunity and reproduction

Additional file 1: Supplementary Figure 1. The extracted sperm-surface proteins. PAGE profiles of... more

Additional file 6 of Buffalo sperm surface proteome profiling reveals an intricate relationship between innate immunity and reproduction

Additional file 6: Supplementary Figure 6. Negative controls for BuBD-129 and 126. Bright field a... more

Deep Metabolomic Profiling Reveals Alterations in Fatty Acid Synthesis and Ketone Body Degradations in Spermatozoa and Seminal Plasma of Astheno-Oligozoospermic Bulls

Frontiers in Veterinary Science, 2022

Male fertility is extremely important in dairy animals because semen from a single bull is used t... more Male fertility is extremely important in dairy animals because semen from a single bull is used to inseminate several thousand females. Asthenozoospermia (reduced sperm motility) and oligozoospermia (reduced sperm concentration) are the two important reasons cited for idiopathic infertility in crossbred bulls; however, the etiology remains elusive. In this study, using a non-targeted liquid chromatography with tandem mass spectrometry-based approach, we carried out a deep metabolomic analysis of spermatozoa and seminal plasma derived from normozoospermic and astheno-oligozoospermic bulls. Using bioinformatics tools, alterations in metabolites and metabolic pathways between normozoospermia and astheno-oligozoospermia were elucidated. A total of 299 and 167 metabolites in spermatozoa and 183 and 147 metabolites in seminal plasma were detected in astheno-oligozoospermic and normozoospermic bulls, respectively. Among the mapped metabolites, 75 sperm metabolites were common to both the g...

Genome Wide Prediction, Mapping and Development of Genomic Resources of Mastitis Associated Genes in Water Buffalo

Frontiers in Veterinary Science, 2021

Water buffalo (Bubalus bubalis) are an important animal resource that contributes milk, meat, lea... more Water buffalo (Bubalus bubalis) are an important animal resource that contributes milk, meat, leather, dairy products, and power for plowing and transport. However, mastitis, a bacterial disease affecting milk production and reproduction efficiency, is most prevalent in populations having intensive selection for higher milk yield, especially where the inbreeding level is also high. Climate change and poor hygiene management practices further complicate the issue. The management of this disease faces major challenges, like antibiotic resistance, maximum residue level, horizontal gene transfer, and limited success in resistance breeding. Bovine mastitis genome wide association studies have had limited success due to breed differences, sample sizes, and minor allele frequency, lowering the power to detect the diseases associated with SNPs. In this work, we focused on the application of targeted gene panels (TGPs) in screening for candidate gene association analysis, and how this approa...

Cellular and Molecular Insights Into the Etiology of Subfertility/Infertility in Crossbred Bulls (Bos taurus × Bos indicus): A Review

Frontiers in Cell and Developmental Biology, 2021

Crossbreeding of indigenous cattle (Bos indicus) with improved (Bos taurus) breeds gained momentu... more Crossbreeding of indigenous cattle (Bos indicus) with improved (Bos taurus) breeds gained momentum and economic relevance in several countries to increase milk production. While production performance of the crossbred offspring is high due to hybrid vigor, they suffer from a high incidence of reproductive problems. Specifically, the crossbred males suffer from serious forms of subfertility/infertility, which can have a significant effect because semen from a single male is used to breed several thousand females. During the last two decades, attempts have been made to understand the probable reasons for infertility in crossbred bulls. Published evidence indicates that testicular cytology indices, hormonal concentrations, sperm phenotypic characteristics and seminal plasma composition were altered in crossbred compared to purebred males. A few recent studies compared crossbred bull semen with purebred bull semen using genomics, transcriptomics, proteomics and metabolomics; molecules p...

RNA-Seq analysis reveals functionally relevant coding and non-coding RNAs in crossbred bull spermatozoa

Animal Reproduction Science, 2020

Transcriptomic Profiling of Buffalo Spermatozoa Reveals Dysregulation of Functionally Relevant mRNAs in Low-Fertile Bulls

Frontiers in Veterinary Science, 2021

Although, it is known that spermatozoa harbor a variety of RNAs that may influence embryonic deve... more Although, it is known that spermatozoa harbor a variety of RNAs that may influence embryonic development, little is understood about sperm transcriptomic differences in relation to fertility, especially in buffaloes. In the present study, we compared the differences in sperm functional attributes and transcriptomic profile between high- and low-fertile buffalo bulls. Sperm membrane and acrosomal integrity were lower (P< 0.05), while protamine deficiency and lipid peroxidation were higher (P< 0.05) in low- compared to high-fertile bulls. Transcriptomic analysis using mRNA microarray technology detected a total of 51,282 transcripts in buffalo spermatozoa, of which 4,050 transcripts were differentially expressed, and 709 transcripts were found to be significantly dysregulated (P< 0.05 and fold change >1) between high- and low-fertile bulls. Majority of the dysregulated transcripts were related to binding activity, transcription, translation, and metabolic processes with pr...

Comparative Transcriptomic Analysis of Spermatozoa From High- and Low-Fertile Crossbred Bulls: Implications for Fertility Prediction

Frontiers in Cell and Developmental Biology, 2021

Crossbred bulls produced by crossing Bos taurus and Bos indicus suffer with high incidence of inf... more Crossbred bulls produced by crossing Bos taurus and Bos indicus suffer with high incidence of infertility/subfertility problems; however, the etiology remains poorly understood. The uncertain predictability and the inability of semen evaluation techniques to maintain constant correlation with fertility demand for alternate methods for bull fertility prediction. Therefore, in this study, the global differential gene expression between high- and low-fertile crossbred bull sperm was assessed using a high-throughput RNA sequencing technique with the aim to identify transcripts associated with crossbred bull fertility. Crossbred bull sperm contained transcripts for 13,563 genes, in which 2,093 were unique to high-fertile and 5,454 were unique to low-fertile bulls. After normalization of data, a total of 776 transcripts were detected, in which 84 and 168 transcripts were unique to high-fertile and low-fertile bulls, respectively. A total of 176 transcripts were upregulated (fold change &g...

Establishment of Repertoire of Placentome-Associated MicroRNAs and Their Appearance in Blood Plasma Could Identify Early Establishment of Pregnancy in Buffalo (Bubalus bubalis)

Frontiers in Cell and Developmental Biology, 2021

Precise early pregnancy diagnosis in dairy animals is of utmost importance for an efficient dairy... more Precise early pregnancy diagnosis in dairy animals is of utmost importance for an efficient dairy production system. Not detecting a dairy animal pregnant sufficiently early after the breeding results to extending the unproductive time of their milk production cycle and causes substantial economic loss for a dairy producer. At present, the most conventional and authentic pregnancy confirmation practice in cows and buffaloes is rectal palpation of the reproductive organs at Days 35–40 after insemination, which sometime leads to considering an animal as false pregnant. Other alternative methods available for early pregnancy diagnosis lack either accuracy or reproducibility or require elaborate instrumentation and laboratory setup not feasible to practice at farmers’ doorstep. The present study was aimed at establishment of the microRNA (miRNA) repertoire of the placentome in buffaloes, which could capture the event of the cross talk between a growing embryo and a dam, through fetal co...