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      Transcriptome analysis to identify candidate genes related to mammary gland development of Bactrian camel ( Camelus bactrianus)

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          Abstract

          Introduction

          The demand for camel milk, which has unique therapeutic properties, is increasing. The mammary gland is the organ in mammals responsible for the production and quality of milk. However, few studies have investigated the genes or pathways related to mammary gland growth and development in Bactrian camels. This study aimed to compare the morphological changes in mammary gland tissue and transcriptome expression profiles between young and adult female Bactrian camels and to explore the potential candidate genes and signaling pathways related to mammary gland development.

          Methods

          Three 2  years-old female camels and three 5  years-old adult female camels were maintained in the same environment. The parenchyma of the mammary gland tissue was sampled from the camels using percutaneous needle biopsy. Morphological changes were observed using hematoxylin-eosin staining. High-throughput RNA sequencing was performed using the Illumina HiSeq platform to analyze changes in the transcriptome between young and adult camels. Functional enrichment, pathway enrichment, and protein–protein interaction networks were also analyzed. Gene expression was verified using quantitative real-time polymerase chain reaction (qRT-PCR).

          Results

          Histomorphological analysis showed that the mammary ducts and mammary epithelial cells in adult female camels were greatly developed and differentiated from those in young camels. Transcriptome analysis showed that 2,851 differentially expressed genes were obtained in the adult camel group compared to the young camel group, of which 1,420 were upregulated, 1,431 were downregulated, and 2,419 encoded proteins. Functional enrichment analysis revealed that the upregulated genes were significantly enriched for 24 pathways, including the Hedgehog signaling pathway which is closely related to mammary gland development. The downregulated genes were significantly enriched for seven pathways, among these the Wnt signaling pathway was significantly related to mammary gland development. The protein–protein interaction network sorted the nodes according to the degree of gene interaction and identified nine candidate genes: PRKAB2, PRKAG3, PLCB4, BTRC, GLI1, WIF1, DKK2, FZD3, and WNT4. The expression of fifteen genes randomly detected by qRT-PCR showed results consistent with those of the transcriptome analysis.

          Discussion

          Preliminary findings indicate that the Hedgehog, Wnt, oxytocin, insulin, and steroid biosynthesis signaling pathways have important effects on mammary gland development in dairy camels. Given the importance of these pathways and the interconnections of the involved genes, the genes in these pathways should be considered potential candidate genes. This study provides a theoretical basis for elucidating the molecular mechanisms associated with mammary gland development and milk production in Bactrian camels.

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          Most cited references57

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          STRING v11: protein–protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets

          Abstract Proteins and their functional interactions form the backbone of the cellular machinery. Their connectivity network needs to be considered for the full understanding of biological phenomena, but the available information on protein–protein associations is incomplete and exhibits varying levels of annotation granularity and reliability. The STRING database aims to collect, score and integrate all publicly available sources of protein–protein interaction information, and to complement these with computational predictions. Its goal is to achieve a comprehensive and objective global network, including direct (physical) as well as indirect (functional) interactions. The latest version of STRING (11.0) more than doubles the number of organisms it covers, to 5090. The most important new feature is an option to upload entire, genome-wide datasets as input, allowing users to visualize subsets as interaction networks and to perform gene-set enrichment analysis on the entire input. For the enrichment analysis, STRING implements well-known classification systems such as Gene Ontology and KEGG, but also offers additional, new classification systems based on high-throughput text-mining as well as on a hierarchical clustering of the association network itself. The STRING resource is available online at https://string-db.org/.
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            A survey of best practices for RNA-seq data analysis

            RNA-sequencing (RNA-seq) has a wide variety of applications, but no single analysis pipeline can be used in all cases. We review all of the major steps in RNA-seq data analysis, including experimental design, quality control, read alignment, quantification of gene and transcript levels, visualization, differential gene expression, alternative splicing, functional analysis, gene fusion detection and eQTL mapping. We highlight the challenges associated with each step. We discuss the analysis of small RNAs and the integration of RNA-seq with other functional genomics techniques. Finally, we discuss the outlook for novel technologies that are changing the state of the art in transcriptomics. Electronic supplementary material The online version of this article (doi:10.1186/s13059-016-0881-8) contains supplementary material, which is available to authorized users.
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              Mammary development in the embryo and adult: a journey of morphogenesis and commitment.

              Mammary gland development occurs through distinctive stages throughout embryonic and pubertal development and reproductive life. At each stage, different signals are required to induce changes in both the epithelium and the surrounding mesenchyme/stroma. Recent studies have provided new insights into the origin, specification and fate of mammary stem and progenitor cells and into how the differentiated lineages that comprise the functional mammary gland are determined. The development of new tools and culture techniques has also enabled the factors that influence branching morphogenesis in the embryonic and pubertal gland to be identified. A surprising recent discovery has been that mammary epithelial cells commit to differentiated lineages using the same signalling pathways that regulate lineage determination in T helper cells.
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                Author and article information

                Contributors
                Journal
                Front Vet Sci
                Front Vet Sci
                Front. Vet. Sci.
                Frontiers in Veterinary Science
                Frontiers Media S.A.
                2297-1769
                05 June 2023
                2023
                : 10
                : 1196950
                Affiliations
                [1] 1Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University , Ürümqi, China
                [2] 2Xinjiang Camel Industry Engineering Technology Research Center , Ürümqi, China
                [3] 3College of Veterinary Medicine, Xinjiang Agricultural University , Ürümqi, China
                [4] 4Xinjiang Altai Regional Animal Husbandry Veterinary Station , Altay, China
                [5] 5Bactrian Camel Academy of Xinjiang, Wangyuan Camel Milk Limited Company , Altay, China
                Author notes

                Edited by: Muhammad Zahoor Khan, University of Agriculture, Dera Ismail Khan, Pakistan

                Reviewed by: Qudrat Ullah, University of Agriculture, Dera Ismail Khan, Pakistan; Lei Liu, Chinese Academy of Agricultural Sciences, China

                *Correspondence: Jie Yang, yangjie234@ 123456xju.edu.cn
                Article
                10.3389/fvets.2023.1196950
                10277799
                37342620
                d111aa5c-498b-4cc5-8aee-f53fbd25fb34
                Copyright © 2023 Yao, Liang, Dou, Zhao, Ma, Hao, Yan, Wang, Wu, Chen and Yang.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 30 March 2023
                : 19 May 2023
                Page count
                Figures: 7, Tables: 2, Equations: 0, References: 57, Pages: 12, Words: 7108
                Funding
                Funded by: Research and Development Program in Xinjiang Uygur Autonomous Region
                Award ID: 2018B01003
                Funded by: Research and Development Projects of China
                Award ID: 2019YFC1606103
                Funded by: Postgraduate Scientific Research Innovation Program of Xinjiang Uygur Autonomous Region
                Award ID: XJ2019G026
                Categories
                Veterinary Science
                Original Research
                Custom metadata
                Livestock Genomics

                bactrian camel,transcriptome sequencing,mammary gland,development,differentially expressed gene

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