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The Council of the Applied Biotechnology Department, for the month of December 2023

 

The Applied Biotechnology Department Council held its session in the institute's council hall in at 10:00 A.m. on Tuesday, 5/12/2023, Under the chairmanship of Prof. Dr. Muhammad Ahmad Almolakab Belkhirshy, Head of Department, and the membership of the following:

Prof. Dr. Heba Atia Yassa

Prof. Dr. Mohammed Sayed Ahmed Moursy

Dr. Marwa Talaat Kamel El Mahdy

Dr. Ghada Abdel Monsef Mahmoud

Dr. Ahmed Mahmoud Hamdy

Dr. Hesham Youseef Awad Darwish

Epigenetic mechanism of Gtl2-miRNAs causes the primitive sheep characteristics found in purebred Merino sheep

Research Abstract

Background

It is not uncommon for some individuals to retain certain primitive characteristics even after domestication or long-term intensive selection. Wild ancestors or original varieties of animals typically possess strong adaptability to environmental preservation, a trait that is often lacking in highly artificially selected populations. In the case of the Merino population, a world-renowned fine wool sheep breed, a phenotype with primitive coarse wool characteristic has re-emerged. It is currently unclear whether this characteristic is detrimental to the production of fine wool or whether it is linked to the adaptability of sheep. The underlying genetic/epigenetic mechanisms behind this trait are also poorly understood.

Results

This study identified lambs with an ancestral-like coarse (ALC) wool type that emerged during the purebred breeding of Merino fine wool sheep. The presence of this primitive sheep characteristic resulted in better environmental adaptability in lambs, as well as improved fine wool yield in adulthood. Reciprocal cross experiments revealed that the ALC phenotype exhibited maternal genetic characteristics. Transcriptomic SNP analysis indicated that the ALC phenotype was localized to the imprinted Gtl2-miRNAs locus, and a significant correlation was found between the ALC wool type and a newly identified short Interstitial Telomeric Sequences (s-ITSs) at this locus. We further confirmed that a novel 38-nt small RNA transcribed from these s-ITSs, in combination with the previously reported 22-nt small RNAs cluster from the Gtl2-miRNAs locus, synergistically inhibited PI3K/AKT/Metabolic/Oxidative stress and subsequent apoptotic pathways in wool follicle stem cells, resulting in the ALC wool type. The necessity of Gtl2-miRNAs in controlling primary hair follicle morphogenesis, as well as the wool follicle type for ALC wool lambs, was verified using intergenic differentially methylated region-knockout mice.

Conclusion

The ALC wool type of Merino sheep, which does not reduce wool quality but increases yield and adaptability, is regulated by epigenetic mechanisms in the imprinted Gtl2-miRNAs region on sheep chromosome 18, with the maternally expressed imprinted gene responsible for the ALC phenotype. This study highlights the significance of epigenetic regulation during embryonic and juvenile stages and emphasizes the advantages of early adaptation breeding for maternal parents in enhancing the overall performance of their offspring.

Research Authors
Jiankui Wang, Guoying Hua, Jianfei Chen, Kai Cui, Zu Yang, Deping Han, Xue Yang, Xianggui Dong, Yuhao Ma, Ganxian Cai, Yuanyuan Zhang, Jinnan Li, Yurong Tai, Lai Da, Xinhai Li, Lina Ma, Qing Ma, Rui Li, Jianbin Liu, Hesham YA Darwish, Keliang Wu, Weiheng
Research Date
Research Journal
Cell & Bioscience
Research Member
Research Pages
190
Research Publisher
BioMed Central
Research Vol
13
Research Website
https://link.springer.com/article/10.1186/s13578-023-01142-z
Research Year
2023

The Immune Barrier of Porcine Uterine Mucosa Differs Dramatically at Proliferative and Secretory Phases and Could Be Positively Modulated by Colonizing Microbiota

Research Abstract

Endometrial immune response is highly associated with the homeostatic balance of the uterus and embryo development; however, the underlying molecular regulatory mechanisms are not fully elucidated. Herein, the porcine endometrium showed significant variation in mucosal immunity in proliferative and secretory phases by single-cell RNA sequencing. The loose arrangement and high motility of the uterine epithelium in the proliferative phase gave opportunities for epithelial cells and dendritic cells to cross talk with colonizing microbial community, guiding lymphocyte migration into the mucosal and glandular epithelium. The migrating lymphocytes were primarily NK and CD8+ T cells, which were robustly modulated by the chemokine signaling. In the secretory phase, the significantly strengthened mechanical mucosal barrier and increased immunoglobulin A alleviated the migration of lymphocytes into the epithelium when the neuro-modulation, mineral uptake, and amino acid metabolism were strongly upregulated. The noticeably increased intraepithelial lymphocytes were positively modulated by the bacteria in the uterine cavity. Our findings illustrated that significant mucosal immunity variation in the endometrium in the proliferative and secretory phases was closely related to intraepithelial lymphocyte migration, which could be modulated by the colonizing bacteria after cross talk with epithelial cells with higher expressions of chemokine.

Research Authors
Deping Han, Peng Sun, Yanxin Hu, Jing Wang, Guoying Hua, Jianfei Chen, Chuyun Shao, Fan Tian, Hesham YA Darwish, Yurong Tai, Xue Yang, Jianyu Chang, Yunfei Ma
Research Date
Research Journal
Frontiers in Immunology
Research Member
Research Pages
750808
Research Publisher
Frontiers Media SA
Research Vol
12
Research Website
https://www.frontiersin.org/articles/10.3389/fimmu.2021.750808/full
Research Year
2021

Genome-wide association study and a post replication analysis revealed a promising genomic region and candidate genes for chicken eggshell blueness

Research Abstract

The eggshell blueness is an interesting object for chicken genetic studies and blue-shelled chicken industry, especially after the discovery of the causative mutation of chicken blue eggshell. In the present study, genome wide association study (GWAS) was conducted in Chinese Dongxiang blue-shelled chicken underlying four traits of blue eggshell pigments: quantity of biliverdin (QB), quantity of protoporphyrin (QP), quantity of total pigment (QT), and color density trait (CD). A total of 139 individuals were randomly collected for GWAS. We detected two SNPs in genome-wise significance and 35 in suggestive significance, 24 out of the 37 SNP were located either within intron/exon or near 15 genes in a range of ~1.17 Mb on GGA21. For further confirmation of the identified SNP loci by GWAS, the follow-up replication studies were performed in two populations. A total of 146 individuals of the second generation derived from the former GWAS population, as well as 280 individuals from an alternative independent population were employed for genotyping by MALDI-TOF MS in a genotype-phenotype association study. Eighteen SNPs evenly distributed on the GGA21 significant region were successfully genotyped in the two populations, of which 4 and 6 SNP loci were shown significantly associated with QB, QT and QP in the two repeat populations, respectively. Further, the SNPs were narrowed down to a region of ~ 653.819 Kb on GGA21 that harbors five candidate genes: AJAP1TNFRSF9C1ORF174CAMTA1, and CEP104. Shell gland of chickens laying dark and light blue eggshell was chosen for detection of mRNA expression of the five candidate genes. The results showed differential expression levels of these genes in the two groups. The specific function of these genes has not yet been defined clearly in chickens and further in-depth studies are needed to explore the new functional role in chicken eggshell blueness.

Research Authors
Hesham YA Darwish, Seyed Benyamin Dalirsefat, Xianggui Dong, Guoying Hua, Jianfei Chen, Yuanyuan Zhang, Jianxiong Li, Jiansheng Xu, Junying Li, Xuemei Deng, Changxin Wu
Research Date
Research Journal
Plos one
Research Member
Research Pages
e0209181
Research Publisher
Public Library of Science
Research Vol
14
Research Website
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0209181
Research Year
2019

Crushing corn pollen grains increased diet digestibility and hemolymph protein content while decreasing honey bee consumption

Research Abstract

A honey bee colony’s ability to grow and develop is dependent on adequate nutrition. Bees collect pollen from flowers as a source of protein, fat, vitamins, and minerals. The crude protein content of corn pollen is considered low, around 15%; however, bees frequently visit the male flowers of the tassels for pollen. In this study, we aimed for the first time to improve the nutritious value of corn pollen by mechanically crushing its external pollen wall. We then compared the effect of feeding crushed vs. non-crushed corn pollen grains on honey bee diet consumption, digestibility, hemolymph protein content, hypopharyngeal gland (HPG) size, and thorax weight under laboratory conditions. We found that crushing corn pollen grains increased diet digestibility and hemolymph protein content while decreasing honey bee pollen consumption (− 39.88%). Crushing pollen however had no effect on HPG size or thorax weight. These findings may be beneficial to beekeepers in areas where corn monoculture is prevalent. The effect of crushed corn pollen on larval development and growth, as well as colony development and vitality, should be investigated in future studies.

Research Authors
Eslam M Omar, Hesham YA Darwish, Ali A Othman, Hesham R El-Seedi, Yahya Al Naggar
Research Date
Research Journal
Apidologie
Research Member
Research Pages
52
Research Publisher
Springer Paris
Research Vol
53
Research Website
https://link.springer.com/article/10.1007/s13592-022-00963-0
Research Year
2022

Molecular cloning and characterization of the endothelin 3 gene in black bone sheep

Research Abstract

Background

Black bone sheep was first discovered in Yunnan province of China in 1970, with unique black pigmentation on the body and internal organs. Endothelin 3 (EDN3) has been known as a key gene causing hyperpigmentation in black bone chicken, the Silky fowl.

Methods

In this study, EDN3 was employed as a candidate gene for regulating black color pigmentation. First, EDN3 was cloned from sheep to obtain the full-length cDNA by using the rapid amplification of cDNA ends (RACE). Genomic EDN3 was screened and a total of thirty predicted single nucleotide polymorphisms (SNPs) were genotyped for allele and genotype frequency analysis in a case-control study involving two black bone sheep populations. Genomic copy number analysis of EDN3 in sheep was conducted to measure the variation in copy number. EDN3 expression levels were observed among the groups in adult liver, lymph node, and kidney tissues, as well as embryo kidney samples. Also, among the tissues of black bone and non-black bone sheep.

Results

The size of the full-length cDNA was 1,578 bp, which included 426 bp of 5′-untranslated region (5′-UTR), an open reading frame (ORF) of 639 bp encoding a protein of 212 amino acids, and a 3′-UTR of 513 bp. Genotype and allele frequencies of all the discovered SNPs were found insignificantly different in black bone and non-black bone sheep (P > 0.05). Genomic copy number analysis of EDN3 in sheep revealed no significant difference between the two sheep groups. No significant variations were found in the adult liver and kidney embryo samples. However, the expression in lymph node and kidney tissue was significantly higher in black bone sheep than that in non-black bone sheep (P < 0.05). Significant variations in the EDN3 expression levels were observed among the tissues of non-black bone sheep.

Conclusions

The findings of the present study indicate that unlike in Silky chickens, EDN3 is not responsible for hyperpigmentation but may play a key functional role in immune and excretory systems of black bone sheep.

Research Authors
Hesham YA Darwish, Yuanyuan Zhang, Kai Cui, Zu Yang, Deping Han, Xianggui Dong, Huaming Mao, Weidong Deng, Xuemei Deng
Research Date
Research Journal
Journal of Animal Science and Biotechnology
Research Member
Research Pages
1-11
Research Publisher
BioMed Central
Research Vol
9
Research Website
https://link.springer.com/article/10.1186/s40104-018-0272-y#Abs1
Research Year
2018

Molecular cloning and characterization of the β-catenin gene from fine-wool sheep

Research Abstract

β-Catenin is an evolutionarily conserved molecule that functions as a crucial effector in both cell-to-cell adhesion and Wnt signaling. To gain a better understanding of its role in the development of hair follicles, we cloned the cDNA sequence of the β-catenin gene from the skin of Aohan fine-wool sheep and performed a variety of bioinformatics analyses. We obtained the full-length sequence, which was 4573-bp long and contained a 2346-bp open reading frame encoding a protein of 781 amino acids. The protein had a predicted molecular weight of 85.4 kDa and a theoretical isoelectric point of 5.57. Domain architecture analysis of the β-catenin protein revealed an armadillo repeat region, which is a common feature of β-catenin in other species. The ovine β-catenin gene shares 97.91%, 94.25%, 94.59%, 83.89%, and 89.39% sequence identity with its homologs in Bos taurus, Homo sapiens, Sus scrofa, Gallus gallus, and Mus musculus, respectively, while the amino acid sequence is more than 99% identical with each of these species. The expression of β-catenin mRNA was detected in the heart, liver, spleen, lung, kidney, skin, muscle, and adipose tissue. Expression levels were maximal in the lung and minimal in the muscle, and the difference in expression in these tissues was significant (P < 0.01). Western blot analysis revealed the presence of the β-catenin protein in all tissues examined; expression was lowest in the skin and adipose tissues.

Research Authors
Kai Cui, Zu Yang, Hesham Darwish, Yuanyuan Zhang, Yaqiong Ge, Xiyue Zhang, Rongni Li, Xuemei Deng
Research Date
Research Journal
Gene
Research Member
Research Pages
277-282
Research Publisher
Elsevier
Research Vol
546
Research Website
https://www.sciencedirect.com/science/article/abs/pii/S0378111914006453
Research Year
2014
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