Serial frequent or multiple Tru-cut® testicular biopsies in rams enable assessment of histological characteristics or transcriptional profiles, with no acute or chronic adverse effects

IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY
A.G.R. Pupulim , P.Z. Rattes , H.D. Mogollón García , J.C. Carvalho , K.Z. Uzae , G.C. Ribeiro , G. Rizzoto , R. Denadai , G.P. Nogueira , F.M.N. Navolar , G.W. Di Santis , S.G. Nunes , A.C.S. Castilho , J.P. Kastelic , J.C.P. Ferreira
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引用次数: 0

Abstract

This study aimed to evaluate the feasibility of performing multiple testicular biopsies in rams using Tru-cut® needles, assessing histological structure, gene expression, and potential complications such as effects on semen quality, testicular blood flow, and ultrasonographic echotexture. In Exp. 1, six mature rams underwent testicular biopsies at intervals (0, 3, 6, 12, 24, and 48 h) using a 16 G Tru-cut® needle, with alternating testes for each collection. Benzathine benzylpenicillin and flunixin meglumine were administered for infection and inflammation control. Local anesthesia and post-biopsy care included lidocaine, digital pressure, and ice application. Testicular samples were analyzed for gene expression related to inflammation, oxidative stress, and steroidogenesis. Semen quality was assessed pre-biopsy and 28 days post-biopsy. Ultrasonographic evaluations of the scrotum and testes were conducted before biopsies and on days 5, 9, 13, 17, and 21 post-biopsies. In Exp. 2, a second group of six mature rams underwent biopsies using 14 G needles, with two samples taken from each testis. Samples were histologically examined for structural preservation. Scrotal skin temperature was measured using infrared thermography, and testicular blood flow was assessed via color Doppler ultrasonography, with measurements taken before and on days 1, 2, 4, 6, 8, 10, 25, 50, 75, and 100 post-biopsies. Semen collection followed the same schedule as in Exp. 1. In Exp. 3, biopsies were performed on different testicular regions (upper, middle, lower) using 12 G, 14 G, and 16 G needles to compare structural preservation. Samples were histologically analyzed. No clinical signs of injury, inflammation, or fluid accumulation were observed. Scrotal pain, increased temperature, swelling, and bleeding were absent, and behavioral signs indicative of pain were not detected. Gene expression remained unchanged, and no significant alterations in seminal characteristics or testicular echogenicity were observed. A slight increase in resistivity and pulsatility indices was noted in Exp. 2. Biopsies with 14 G and 16 G needles resulted in structural disruptions, while 12 G needles better preserved testicular parenchyma. Multiple testicular biopsies using Tru-cut® needles did not cause significant morphological changes, alter transcriptional profiles, or affect semen or ultrasonographic characteristics, demonstrating that this method is viable for monitoring acute molecular changes in the testes.
对公羊进行连续、频繁或多次的 Tru-cut® 睾丸活检,可评估组织学特征或转录谱,且无急性或慢性不良影响。
本研究旨在评估使用 Tru-cut® 注射针对公羊进行多次睾丸活检的可行性,评估组织学结构、基因表达以及潜在并发症(如对精液质量、睾丸血流和超声回声纹理的影响)。在实验 1 中,六只成熟公羊使用 16 G Tru-cut® 针每隔一段时间(0、3、6、12、24 和 48 小时)进行睾丸活检,每次采集交替进行。苄星青霉素和氟尼辛葡胺用于控制感染和炎症。局部麻醉和活检后护理包括利多卡因、数字加压和冰敷。对睾丸样本进行了与炎症、氧化应激和类固醇生成相关的基因表达分析。精液质量在活检前和活检后 28 天进行评估。在活组织检查前以及活组织检查后的第 5、9、13、17 和 21 天,对阴囊和睾丸进行了超声波评估。在实验 2 中,第二组的六只成熟公羊使用 14 G 的针头进行了活组织检查,每个睾丸取两个样本。对样本进行组织学检查,以确定其结构是否保存完好。使用红外热成像技术测量阴囊皮肤温度,并通过彩色多普勒超声波检查评估睾丸血流量,在活检前和活检后第 1、2、4、6、8、10、25、50、75 和 100 天进行测量。精液采集时间与实验 1 相同。在实验 3 中,使用 12 G、14 G 和 16 G 的活检针对不同的睾丸区域(上、中、下)进行活检,以比较结构保存情况。对样本进行了组织学分析。没有观察到损伤、炎症或积液的临床症状。阴囊无疼痛、温度升高、肿胀和出血,也未发现表明疼痛的行为体征。基因表达保持不变,精液特征和睾丸回声也没有明显改变。使用 14 G 和 16 G 的活检针进行活检会导致结构破坏,而 12 G 的活检针能更好地保留睾丸实质。使用Tru-cut®针进行多次睾丸活检不会导致明显的形态学变化,也不会改变转录谱,或影响精液或超声波特征,这表明这种方法可用于监测睾丸的急性分子变化。
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来源期刊
Theriogenology
Theriogenology 农林科学-生殖生物学
CiteScore
5.50
自引率
14.30%
发文量
387
审稿时长
72 days
期刊介绍: Theriogenology provides an international forum for researchers, clinicians, and industry professionals in animal reproductive biology. This acclaimed journal publishes articles on a wide range of topics in reproductive and developmental biology, of domestic mammal, avian, and aquatic species as well as wild species which are the object of veterinary care in research or conservation programs.
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