Maternal homozygosity for three independent mutant hecate alleles results in embryos with reduced expression of dorsal organizer genes and defects in the formation of dorsoanterior structures. A positional cloning approach identified all hecate mutations as stop codons affecting the same gene, revealing that hecate encodes the Glutamate receptor interacting protein 2a (Grip2a), a protein containing multiple PDZ domains known to interact with membrane-associated factors including components of the Wnt signaling pathway. We find that grip2a mRNA is localized to the vegetal pole of the oocyte and early embryo, and that during egg activation this mRNA shifts to an off-center vegetal position corresponding to the previously proposed teleost cortical rotation. hecate mutants show defects in the alignment and bundling of microtubules at the vegetal cortex, which result in defects in the asymmetric movement of wnt8a mRNA as well as anchoring of the kinesin-associated cargo adaptor Syntabulin. We also find that, although short-range shifts in vegetal signals are affected in hecate mutant embryos, these mutants exhibit normal long-range, animally directed translocation of cortically injected dorsal beads that occurs in lateral regions of the yolk cortex. Furthermore, we show that such animally-directed movement along the lateral cortex is not restricted to a single arc corresponding to the prospective dorsal region, but occur in multiple meridional arcs even in opposite regions of the embryo. Together, our results reveal a role for Grip2a function in the reorganization and bundling of microtubules at the vegetal cortex to mediate a symmetry-breaking short-range shift corresponding to the teleost cortical rotation. The slight asymmetry achieved by this directed process is subsequently amplified by a general cortical animally-directed transport mechanism that is neither dependent on hecate function nor restricted to the prospective dorsal axis.
The aim of the present study was to investigate the physicochemical and microbiological profile of indigenous sheep’s milk with the potential for the production of artisanal cheese. The data showed the changes in lactation stages (between April and August) up to 2 hours following manual milk production. The quality of raw milk in the production of brined cheese using traditional technology was assessed. The amount of dry matter was21.36% in April and 19.85% in August at the end of lactation. The fat content from April to August was between 8.62% and 8.03%, preserving the values reported. The protein composition of raw milk varied between 5.52 – 6.61%. In the microbiological evaluation of raw sheep’s milk after milking (35.3°C), the total microbial contamination was in the range of 1.3 x 106 to 1.4 x 106 CFU/mL under current conditions. The results obtained in this paper give rise to more in-depth studies on the application of raw sheep’s milk in the production of traditional artisanal cheeses.
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