Genetic Dissection of Transcriptional Regulation in Budding Yeast

Cryopreservation advances capacitation-like changes in ram spermatozoa. These changes are reflected in an increased fertilizing ability compared with fresh spermatozoa, followed by an accelerated decline in fertilizing ability after incubation in vitro or in vivo. Furthermore, frozen-thawed spermatozoa are released earlier than fresh spermatozoa after binding to oviduct cells in vitro, confirming their physiological readiness to participate in fertilization despite their short lifespan. After insemination large numbers of spermatozoa are lost from the female reproductive tract of the ewe via the vagina. Frozen-thawed spermatozoa are expelled faster than fresh spermatozoa. The advanced membrane status of frozen-thawed spermatozoa may provoke their rapid loss and possibly makes them more vulnerable to attack by uterine leucocytes, or by some other mechanism, as a high proportion of spermatozoa lost from the tract are decapitated. The observed destabilization of the membranes of cryopreserved spermatozoa is accompanied by impaired sperm transport, associated with mitochondrial injury, necessitating intrauterine deposition of frozen-thawed semen to obtain satisfactory fertility after artificial insemination. However, the frozen-thawed spermatozoa that can participate in fertilization may contribute to increased embryonic loss by the advancement of cleavage or through a direct effect of cryopreservation on the male genome.

/pdf/the-functional-integrity-and-fate-of-cryopreserved-ram-1bvd9lwxez.pdf

The functional integrity and fate of cryopreserved ram spermatozoa in the female tract

Physiology of reproduction in the woman

The domestic water buffalo (Bubalus bubalis) makes a major contribution to the global agricultural economy in the form of milk, meat, hides, and draught power. The global water buffalo population is predominantly found in Asia, and per head of population more people depend upon the buffalo than on any other livestock species. Despite its agricultural importance, there are comparatively fewer genomic and transcriptomic resources available for buffalo than for other livestock species. We have generated a large-scale gene expression atlas covering multiple tissue and cell types from all major organ systems collected from three breeds of riverine water buffalo (Mediterranean, Pandharpuri and Bhadawari) and used the network analysis tool Graphia Professional to identify clusters of genes with similar expression profiles. Alongside similar data, we and others have generated for ruminants as part of the Functional Annotation of Animal Genomes Consortium; this comprehensive transcriptome supports functional annotation and comparative analysis of the water buffalo genome.

/pdf/a-gene-expression-atlas-of-the-domestic-water-buffalo-4sb5up37hg.pdf

A Gene Expression Atlas of the Domestic Water Buffalo (Bubalus bubalis)

The economic importance of genetically improving feed efficiency has been recognized by cattle producers worldwide. It has the potential to considerably reduce costs, minimize environmental impact,...

/pdf/genetic-mechanisms-underlying-feed-utilization-and-3920a3xi9g.pdf

Genetic mechanisms underlying feed utilization and implementation of genomic selection for improved feed efficiency in dairy cattle

Selection for feed efficiency is crucial for overall profitability and sustainability in dairy cattle production. Key regulator genes and genetic markers derived from co-expression networks underlying feed efficiency could be included in the genomic selection of the best cows. The present study identified co-expression networks associated with high and low feed efficiency and their regulator genes in Danish Holstein and Jersey cows. RNA-sequencing data from Holstein and Jersey cows with high and low residual feed intake (RFI) and treated with two diets (low and high concentrate) were used. Approximately 26 million and 25 million pair reads were mapped to bovine reference genome for Jersey and Holstein breed, respectively. Subsequently, the gene count expressions data were analysed using a Weighted Gene Co-expression Network Analysis (WGCNA) approach. Functional enrichment analysis from Ingenuity® Pathway Analysis (IPA®), ClueGO application and STRING of these modules was performed to identify relevant biological pathways and regulatory genes. WGCNA identified two groups of co-expressed genes (modules) significantly associated with RFI and one module significantly associated with diet. In Holstein cows, the salmon module with module trait relationship (MTR) = 0.7 and the top upstream regulators ATP7B were involved in cholesterol biosynthesis, steroid biosynthesis, lipid biosynthesis and fatty acid metabolism. The magenta module has been significantly associated (MTR = 0.51) with the treatment diet involved in the triglyceride homeostasis. In Jersey cows, the lightsteelblue1 (MTR = − 0.57) module controlled by IFNG and IL10RA was involved in the positive regulation of interferon-gamma production, lymphocyte differentiation, natural killer cell-mediated cytotoxicity and primary immunodeficiency. The present study provides new information on the biological functions in liver that are potentially involved in controlling feed efficiency. The hub genes and upstream regulators (ATP7b, IFNG and IL10RA) involved in these functions are potential candidate genes for the development of new biomarkers. However, the hub genes, upstream regulators and pathways involved in the co-expressed networks were different in both breeds. Hence, additional studies are required to investigate and confirm these findings prior to their use as candidate genes.

/pdf/gene-co-expression-networks-from-rna-sequencing-of-dairy-acdgkm5f77.pdf

Gene co-expression networks from RNA sequencing of dairy cattle identifies genes and pathways affecting feed efficiency

The Ovum Pick Up-In vitro Production (OPU-IVP) of embryos is an advanced reproductive technology used in cattle production but the complex biological mechanisms behind IVP outcomes are not fully understood. In this study we sequenced RNA of granulosa cells collected from Holstein cows at oocyte aspiration prior to IVP, to identify candidate genes and biological mechanisms for favourable IVP-related traits in donor cows where IVP was performed separately for each animal. We identified 56 genes significantly associated with IVP scores (BL rate, kinetic and morphology). Among these, BEX2, HEY2, RGN, TNFAIP6 and TXNDC11 were negatively associated while Mx1 and STC1 were positively associated with all IVP scores. Functional analysis highlighted a wide range of biological mechanisms including apoptosis, cell development and proliferation and four key upstream regulators (COX2, IL1, PRL, TRIM24) involved in these mechanisms. We found a range of evidence that good IVP outcome is positively correlated with early follicular atresia. Furthermore we showed that high genetic index bulls can be used in breeding without reducing the IVP performances. These findings can contribute to the development of biomarkers from follicular fluid content and to improving Genomic Selection (GS) methods that utilize functional information in cattle breeding, allowing a widespread large scale application of GS-IVP.

/pdf/identification-of-potential-biomarkers-in-donor-cows-for-in-2iwggby71g.pdf

Identification of potential biomarkers in donor cows for in vitro embryo production by granulosa cell transcriptomics.

A better understanding of the hormonal profile of oestrus synchronised Saanen crossbreds would guide appropriate timing of Fixed Time Artificial Insemination (FTAI). This study was conducted to determine the progesterone hormone (P4) and luteinising hormone (LH) concentrations from the end of oestrus synchronisation treatment until the estimated time of ovulation. The LH peak would help in estimating the time of ovulation and appropriate time for FTAI. Twenty three Saanen crossbred does aged 2 to 4 y were allocated to three treatment groups: i) CIDR for 14 d and PMSG and PG at CIDR removal (CIDR14++; n=8); ii) CIDR for 9 d and PMSG and PG at CIDR removal (CIDR9++; n=8); iii) CIDR for 9 d and only PG at CIDR removal (CIDR9+; n=7) intramuscularly. Oestrus signs were recorded and blood samples were collected every 4 h beginning at 24 h post CIDR removal for 24 h. The serum samples were analyzed for P4 and LH concentration using ELISA kits. The CIDR9++ does had shorter (p<0.05) interval between CIDR removal and onset of oestrus than the other two treatment groups. All protocols showed declining P4 hormone concentration after 24 h of CIDR removal. The CIDR14++ does had highest mean LH concentration at peak (8.03±2.07 mIU/ml), followed by CIDR9+ does (7.32±1.07 mIU/ml); these values were higher (P<0.05) than the mean for the CIDR9++ does (4.94±0.80 mIU/ml). The LH peak occurred at 42.0±1.15, 41.0±1.91, and 43.0±1.00 h after CIDR removal for the CIDR14++, CIDR9++, and CIDR9+ treatments, respectively. The use of the intravaginal CIDR for oestrus synchronisation may be shortened in goats to 9 d while there was no significant difference in the time of occurrence of LH peak.

Progesterone and luteinising hormone profile of Saanen crossbred goats synchronised with CIDR and PMSG.

There is lack of information on the effects of oestrus synchronisation using Controlled Internal Drug Release (CIDR) insert in goats in Malaysia. Understanding the effect of oestrus synchronisation would enable the use of fixed-time artificial insemination (FTAI), which might increase the efficiency of goat production. This study will determine the most suitable oestrus synchronisation protocol and FTAI to be practiced in goat industries in Malaysia. Two experiments were conducted in this study. In the first experiment, 23 Saanen crossbred does (2 to 4 years old) were divided unequally into 3 groups. The first group received CIDR for 14 days, Pregnant Mare Serum Gonadotrophin (PMSG) and cloprostenol at CIDR removal (CIDR14++; n=8); the second group of does received CIDR for 9 days and PMSG and cloprostenol at CIDR removal (CIDR9++; n=8) and the third group received CIDR for 9 days and cloprostenol only at CIDR removal (CIDR9+; n=7). Oestrus signs were recorded and blood samples for progesterone (P4) and luteinising hormone (LH) analysis were collected for 24 h at 4 h interval, starting from 24 h post CIDR removal. Data on oestrus signs, P4 and LH were analysed using one-way ANOVA. Results showed 100 % of does in group CIDR14++ in oestrus, while the other two groups (CIDR9++ and CIDR9+) showed only 57% and 33 %, respectively. The time from CIDR removal until onset of oestrus in CIDR9+ was significantly longer (P 0.05) in all groups. The time of LH peak in group CIDR14+ was significantly delayed (P<0.05) when compared to group CIDR9++. The pregnancy rate of FTAI does was not significantly different (P<0.05). In conclusion, the suitable oestrus synchronisation protocols for goat in Malaysia were CIDR14++ and CIDR9++. It is recommended to use the treatment CIDR9++, since the oestrous cycle can be shortened. Since the pregnancy rate in this study is very low; the suitable time to conduct fixed-time AI cannot be concluded.

Reproductive response of goats to different oestrus synchronisation protocols and their combination with fixed time artificial insemination

There is still a lack of information on estrus synchronization in goats. Understanding the estrus synchronization protocols and the subsequent effects is important to improve the efficiency of assisted reproductive technologies (ARTs) and subsequently would improve the breeding procedures. This study will help in determining the most suitable estrus synchronization protocol and understand better the effect on the sexual behaviour and hormonal effects in goats. A total of 127 Boer does were used and divided into three groups with different duration of CIDR insertion intravaginally either for 14 (two groups) or 9 days (one group). Approximately 0.5 ml Estrumate® (PG) was administered intramuscularly to all groups at CIDR removal, and only groups PMSG14 and PMSG9 were administered with 200IU of Pregnant Mare Serum Gonadotropin (PMSG) intramuscularly. Estrus signs were observed at 4 h intervals and blood samples were collected for progesterone and luteinizing hormone determination. The percentage of does in estrus within 24 to 72 h post CIDR removal was significantly higher (P 0.05) in all groups. The time of the LH peak in the group without the PMSG was significantly delayed (P<0.05) when compared to group 9 days CIDR and administered with PMSG. It is recommended to use the treatment for 9 days CIDR since the estrous cycle can be shortened.

/pdf/study-of-sexual-behaviours-with-different-types-of-estrus-34hhc2xaty.pdf

Suraya Mohamad Salleh

Papers

Gene co-expression networks from RNA sequencing of dairy cattle identifies genes and pathways affecting feed efficiency

Identification of potential biomarkers in donor cows for in vitro embryo production by granulosa cell transcriptomics.

Progesterone and luteinising hormone profile of Saanen crossbred goats synchronised with CIDR and PMSG.

Reproductive response of goats to different oestrus synchronisation protocols and their combination with fixed time artificial insemination

Study of sexual behaviours with different types of estrus synchronization protocols in Boer goats.