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Orthotopic transplantation of neonatal GFP rat ovary as experimental model to study ovarian development and toxicology.

ABSTRACT: The rat is one of the most commonly used experimental animal species in biomedical research. The availability of new research tools in rats could therefore provide considerable advances in the areas where this mammal is extensively used. We report the development of a new green fluorescent protein (GFP) rat strain suitable for organ transplantation and the birth of GFP rats following orthotopic transplantation of neonatal ovaries from this newly developed GFP rat strain to a wild-type Fischer 344 (F344) strain. A new GFP rat strain was developed by backcrossing eGFP Sprague-Dawley (SD-Tg(CAG-EGFP)Cz-004Osb) to wild-type F344 for eight generations. Whole ovaries from postnatal day (PND) 8 or PND 21 GFP rats were transplanted orthotopically to bilaterally ovariectomized wild-type adult females (n=6). All recipients were mated, and three of the five resulting litters contained GFP pups. In the PND 8 group, all recipients cycled regularly and the ovarian morphology appeared normal when collected at 9 months post-transplantation. In the PND 21 group, 60% of the recipients displayed regular estrous cycles at 9 months post-transplantation, but showed reduced ovarian size. This new strain and neonatal orthotopic transplantation could be useful for many biomedical fields including transplantation, development, and reproductive toxicology.

SUBMITTER: Marano JE

PROVIDER: S-EPMC2613850 | biostudies-literature | 2008 Nov-Dec

REPOSITORIES: biostudies-literature

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Orthotopic transplantation of neonatal GFP rat ovary as experimental model to study ovarian development and toxicology.

Marano Jason E JE Sun Dongming D Zama Aparna Mahakali AM Young Wise W Uzumcu Mehmet M

Reproductive toxicology (Elmsford, N.Y.) 20080919 3-4

The rat is one of the most commonly used experimental animal species in biomedical research. The availability of new research tools in rats could therefore provide considerable advances in the areas where this mammal is extensively used. We report the development of a new green fluorescent protein (GFP) rat strain suitable for organ transplantation and the birth of GFP rats following orthotopic transplantation of neonatal ovaries from this newly developed GFP rat strain to a wild-type Fischer 34 ...[more]

PMID: 18848623

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Project description:Clinically, peripheral nerve reconstructions in neonates are most frequently applied in brachial plexus birth injuries. Most surgical concepts, however, have investigated nerve reconstructions in adult animal models. The immature neuromuscular system reacts differently to the effects of nerve lesion and surgery and is poorly investigated due to the lack of reliable experimental models. Here, we describe an experimental forelimb model in the neonatal rat, to study these effects on both the peripheral and central nervous systems. Within 24 hours after birth, three groups were prepared: In the nerve transfer group, a lesion of the musculocutaneous nerve was reconstructed by selectively transferring the ulnar nerve. In the negative control group, the musculocutaneous nerve was divided and not reconstructed and in the positive control group, a sham surgery was performed. The animal´s ability to adapt to nerve lesions and progressive improvement over time were depict by the Bertelli test, which observes the development of grooming. Twelve weeks postoperatively, animals were fully matured and the nerve transfer successfully reinnervated their target muscles, which was indicated by muscle force, muscle weight, and cross sectional area evaluation. On the contrary, no spontaneous regeneration was found in the negative control group. In the positive control group, reference values were established. Retrograde labeling indicated that the motoneuron pool of the ulnar nerve was reduced following nerve transfer. Due to this post-axotomy motoneuron death, a diminished amount of motoneurons reinnervated the biceps muscle in the nerve transfer group, when compared to the native motoneuron pool of the musculocutaneous nerve. These findings indicate that the immature neuromuscular system behaves profoundly different than similar lesions in adult rats and explains reduced muscle force. Ultimately, pathophysiologic adaptations are inevitable. The maturing neuromuscular system, however, utilizes neonatal capacity of regeneration and seizes a variety of compensation mechanism to restore a functional extremity. The above described neonatal rat model demonstrates a constant anatomy, suitable for nerve transfers and allows all standard neuromuscular analyses. Hence, detailed investigations on the pathophysiological changes and subsequent effects of trauma on the various levels within the neuromuscular system as well as neural reorganization of the neonatal rat may be elucidated. This study was approved by the Ethics Committee of the Medical University of Vienna and the Austrian Ministry for Research and Science (BMWF-66.009/0187-WF/V/3b/2015) on March 20, 2015.

Project description:The lung is, more than other solid organs, susceptible for ischemia reperfusion injury after orthotopic transplantation. Corticosteroids are known to potently suppress pro-inflammatory processes when given in the post-operative setting or during rejection episodes. Whereas their use has been approved for these clinical indications, there is no study investigating its potential as a preservation additive in preventing vascular damage already in the phase of ischemia. To investigate these effects we performed orthotopic lung transplantations (LTX) in the rat. Prednisolone was either added to the perfusion solution for lung preservation or omitted and rats were followed for 48 hours after LTX. Prednisolone preconditioning significantly increased survival and diminished reperfusion edema. Hypoxia induced vasoactive cytokines such as VEGF were reduced. Markers of leukocyte invasiveness like matrix metalloprotease (MMP)-2, or common pro-inflammatory molecules like the CXCR4 receptor or the chemokine (C-C motif) ligand (CCL)-2 were downregulated by prednisolone. Neutrophil recruitment to the grafts was only increased in Perfadex treated lungs. Together with this, prednisolone treated animals displayed significantly reduced lung protein levels of neutrophil chemoattractants like CINC-1, CINC-2α/β and LIX and upregulated tissue inhibitor of matrix metalloproteinase (TIMP)-1. Interestingly, lung macrophage invasion was increased in both, Perfadex and prednisolone treated grafts, as measured by MMP-12 or RM4. Markers of anti-inflammatory macrophage transdifferentiation like MRC-1, IL-13, IL-4 and CD163, significantly correlated with prednisolone treatment. These observations lead to the conclusion that prednisolone as an additive to the perfusion solution protects from hypoxia triggered danger signals already in the phase of ischemia and thus reduces graft edema in the phase of reperfusion. Additionally, prednisolone preconditioning might also lead to macrophage polarization as a beneficial long-term effect.

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Orthotopic transplantation of neonatal GFP rat ovary as experimental model to study ovarian development and toxicology.

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Orthotopic transplantation of neonatal GFP rat ovary as experimental model to study ovarian development and toxicology.

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