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Since the domestication of cattle more than 10,000 years ago, cattle have been critical in the shift of human society from nomadic hunter-gatherers to sedentary farming communities across most of Europe, Asia, and Africa. Although our understanding of ancestral population relationships is limited, cattle domestication is thought to have occurred on two or three occasions, giving rise to the taurine (B. taurus) and indicine (B. indicus) species, which share a common ancestor with the aurochs (B. primigenius) 250,000 years ago. Indicine and taurine cattle were domesticated in the Indus Valley and the Fertile Crescent, respectively; however, a third domestication event for taurine has been hypothesized in Egypt's Western Desert. Because of their recent split, African indicine cattle share a lot of genetic variation with Asian indicine cattle, as well as with African taurine cattle through gene flow. Although further research is needed to detangle the complicated human-mediated dispersion patterns of domestic cattle, scenarios involving unidirectional or bidirectional migratory events between European taurine and Asian indicine cattle are also feasible. As a result, our research contributes to a better understanding of the impact of previous demographic history on present cow genetic variation, laying the groundwork for future research into alternate migration pathways for early domestic populations. [ABSTRACT FROM AUTHOR]

Simple Summary: Aquaporins are membrane channels that allow for the movement of water and solutes in cells. They have been reported to play crucial roles in mammalian early development and cryopreservation processes. However, there are few studies focused on the characterization of aquaporins in cumulus oocyte and embryo complexes of cattle. Moreover, no studies have been carried out on Brahman, Holstein, Gir and Romosinuano, important bovine breeds in milk and meat production. Therefore, the objective of this study was to evaluate their presence, transcript level and possible functions in the cumulus oocyte complex of the Brahman, Holstein, Gir and Romosinuano breeds and in embryos from five bovine crosses. Aquaporins 1–12 were found in both cumulus oocyte complexes and embryos, and we found possible parental effects on the expression of aquaporins 6 and 12b in cumulus oocyte complexes and aquaporins 4, 8 and 9 in embryos. This allows one to evidence possible functions in the early development of the Brahman, Holstein, Gir and Romosinuano bovine breeds. Aquaporins (AQPs) are proteins with various functions related to proper cell function and early development in mammals. The aim of this study was to evaluate the presence of AQPs and determine their mRNA levels in the cumulus oocyte complex (COC) of four bovine breeds and in blastocysts of five bovine crosses. Grade I, II and III COCs were collected by ovum pick up from non-lactating heifers of the Brahaman, Holstein, Gir and Romosinuano breeds. Embryos were produced in vitro up to the blastocyst stage of the bovine ♀Gir × ♂Holstein, ♀Holstein × ♂Gir, ♀Brahman × ♂Holstein, ♀Holstein × ♂Brahman, and ♀Romosinuano × ♂Holstein crosses. mRNA expression of AQP1-AQP12b was estimated in COC and embryos by real-time-PCR. The presence of the twelve AQPs in the COCs and bovine embryos was established. Additionally, significant differences were determined in the expression of AQP6 and AQP12b in COCs, as well as in transcripts levels of AQP4, AQP8 and AQP9 from bovine embryos. Gene expression of AQPs in COCs and bovine embryos is consistent with the previously described biological functions. This is the first report of AQPs in COC of Gir, Brahman, Holstein and Romosinuano and embryos of five crossbreeds between Bos indicus and B. taurus. [ABSTRACT FROM AUTHOR]

The disassembly of microtubules can generate force and drive intracellular motility. During mitosis, for example, chromosomes remain persistently attached via kinetochores to the tips of disassembling microtubules, which pull the sister chromatids apart. According to the conformational wave hypothesis, such force generation requires that protofilaments curl outward from the disassembling tips to exert pulling force directly on kinetochores. Rigorously testing this idea will require modifying the mechanical and energetic properties of curling protofilaments, but no way to do so has yet been described. Here, by direct measurement of working strokes generated in vitro by curling protofilaments, we show that their mechanical energy output can be increased by adding magnesium, and that yeast microtubules generate larger and more energetic working strokes than bovine microtubules. Both the magnesium and species-dependent increases in work output can be explained by lengthening the protofilament curls, without any change in their bending stiffness or intrinsic curvature. These observations demonstrate how work output from curling protofilaments can be tuned and suggest evolutionary conservation of the amount of curvature strain energy stored in the microtubule lattice.

Mammalian sperm rolling around their longitudinal axes is a long-observed component of motility, but its function in the fertilization process, and more specifically in sperm migration within the female reproductive tract, remains elusive. While investigating bovine sperm motion under simple shear flow and in a quiescent microfluidic reservoir and developing theoretical and computational models, we found that rolling regulates sperm navigation in response to the rheological properties of the sperm environment. In other words, rolling enables a sperm to swim progressively even if the flagellum beats asymmetrically. Therefore, a rolling sperm swims stably along the nearby walls (wall-dependent navigation) and efficiently upstream under an external fluid flow (rheotaxis). By contrast, an increase in ambient viscosity and viscoelasticity suppresses rolling, consequently, non-rolling sperm are less susceptible to nearby walls and external fluid flow and swim in two-dimensional diffusive circular paths (surface exploration). This surface exploration mode of swimming is caused by the intrinsic asymmetry in flagellar beating such that the curvature of a sperm’s circular path is proportional to the level of asymmetry. We found that the suppression of rolling is reversible and occurs in sperm with lower asymmetry in their beating pattern at higher ambient viscosity and viscoelasticity. Consequently, the rolling component of motility may function as a regulatory tool allowing sperm to navigate according to the rheological properties of the functional region within the female reproductive tract.

During mitosis, kinetochore-attached microtubules form bundles (k-fibers) in which many filaments grow and shorten in near-perfect unison to align and segregate each chromosome. However, individual microtubules grow at intrinsically variable rates, which must be tightly regulated for a k-fiber to behave as a single unit. This exquisite coordination might be achieved biochemically, via selective binding of polymerases and depolymerases, or mechanically, because k-fiber microtubules are coupled through a shared load that influences their growth. Here, we use a novel dual laser trap assay to show that microtubule pairs growing in vitro are coordinated by mechanical coupling. Kinetic analyses show that microtubule growth is interrupted by stochastic, force-dependent pauses and indicate persistent heterogeneity in growth speed during non-pauses. A simple model incorporating both force-dependent pausing and persistent growth speed heterogeneity explains the measured coordination of microtubule pairs without any free fit parameters. Our findings illustrate how microtubule growth may be synchronized during mitosis and provide a basis for modeling k-fiber bundles with three or more microtubules, as found in many eukaryotes., Competing Interests: BL, KK, YL, CN, SB, CA No competing interests declared, (© 2023, Leeds et al.)

Abstract: Embryo transfer using cryopreserved B. indicus embryos results in low pregnancy rates. The low viability of B. indicus embryos is likely a result of their low freezability relative to embryos from other species, notably B. taurus. Freezability is probably related to the quantity of lipid droplets in the embryonic cells. However, the mechanism underlying this association is unknown. Using multiphoton and transmission electron microscopy, the proportion, volume, morphology and composition of lipid droplets in fresh and cryopreserved B. indicus and B. taurus embryos were studied. B. indicus embryos have more droplets than B. taurus and a greater percentage of lipid droplets in clusters (34% vs. 24%; p < 0.05). Also, B. indicus individual droplets are of greater volume than those of B. taurus (47 μm3 vs. 34 μm3; p < 0.05). After freezing, the percentage of clusters decreased in B. indicus (11% vs. 5%; p < 0.05), while in B. taurus they increased (2% vs. 7%; p < 0.05). Freezing influenced the lipid droplet morphology, increasing the number of cracked droplets in B. indicus embryos (33% fresh vs. 62% frozen; p < 0.05), but not in B. taurus embryos (35% fresh vs. 34% frozen; ns). Regarding composition, saturated lipid droplets prevail in B. indicus embryos, contrary to B. taurus embryos. We observed differences in lipid droplet organization, proportion, volume, morphology and composition between B. indicus and B. taurus embryos which are altered in opposite directions following freezing. [ABSTRACT FROM AUTHOR]

Due to evolutionary divergence, cattle (taurine, and indicine) and buffalo are speculated to have different responses to heat stress condition. Variation in candidate genes associated with a heat-shock response may provide an insight into the dissimilarity and suggest targets for intervention. The present work was undertaken to characterize one of the inducible heat shock protein genes promoter and coding regions in diverse breeds of Indian zebu cattle and buffaloes. The genomic DNA from a panel of 117 unrelated animals representing 14 diversified native cattle breeds and 6 buffalo breeds were utilized to determine the complete sequence and gene diversity of HSP70.1 gene. The coding region of HSP70.1 gene in Indian zebu cattle, Bos taurus and buffalo was similar in length (1,926 bp) encoding a HSP70 protein of 641 amino acids with a calculated molecular weight (Mw) of 70.26 kDa. However buffalo had a longer 5' and 3' untranslated region (UTR) of 204 and 293 nucleotides respectively, in comparison to Indian zebu cattle and Bos taurus wherein length of 5' and 3'-UTR was 172 and 286 nucleotides, respectively. The increased length of buffalo HSP70.1 gene compared to indicine and taurine gene was due to two insertions each in 5' and 3'-UTR. Comparative sequence analysis of cattle (taurine and indicine) and buffalo HSP70.1 gene revealed a total of 54 gene variations (50 SNPs and 4 INDELs) among the three species in the HSP70.1 gene. The minor allele frequencies of these nucleotide variations varied from 0.03 to 0.5 with an average of 0.26. Among the 14 B. indicus cattle breeds studied, a total of 19 polymorphic sites were identified: 4 in the 5'-UTR and 15 in the coding region (of these 2 were non-synonymous). Analysis among buffalo breeds revealed 15 SNPs throughout the gene: 6 at the 5' flanking region and 9 in the coding region. In bubaline 5'-UTR, 2 additional putative transcription factor binding sites (Elk-1 and C-Re1) were identified, other than three common sites (CP2, HSE and Pax-4) observed across all the analyzed animals. No polymorphism was found within the 3'-UTR of Indian cattle or buffalo as it was found to be monomorphic. The promoter sequences generated in 117 individuals showed a rich array of sequence elements known to be involved in transcription regulation. A total of 11 nucleotide changes were observed in the promoter sequence across the analyzed species, 3 of these changes were located within the potential transcription factor binding domains. We also identified 4 microsatellite markers within the buffalo HSP70.1 gene and 3 microsatellites within bovine HSP70.1. The present study identified several distinct changes across indicine, taurine and bubaline HSP70.1 genes that could further be evaluated as molecular markers for thermotolerance., (Copyright © 2013 Elsevier B.V. All rights reserved.)