Genetic targeting of adult Renshaw cells using a Calbindin 1 destabilized Cre allele for intersection with Parvalbumin or Engrailed1
Abstract Renshaw cells (RCs) are one of the most studied spinal interneurons; however, their roles in motor control remain enigmatic in part due to the lack of experimental models to interfere with RC function, specifically in adults. To overcome this limitation, we leveraged the distinct temporal r...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2021-10-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-99333-6 |
| _version_ | 1818851587426091008 |
|---|---|
| author | Alicia R. Lane Indeara C. Cogdell Thomas M. Jessell Jay B. Bikoff Francisco J. Alvarez |
| author_facet | Alicia R. Lane Indeara C. Cogdell Thomas M. Jessell Jay B. Bikoff Francisco J. Alvarez |
| author_sort | Alicia R. Lane |
| collection | DOAJ |
| description | Abstract Renshaw cells (RCs) are one of the most studied spinal interneurons; however, their roles in motor control remain enigmatic in part due to the lack of experimental models to interfere with RC function, specifically in adults. To overcome this limitation, we leveraged the distinct temporal regulation of Calbindin (Calb1) expression in RCs to create genetic models for timed RC manipulation. We used a Calb1 allele expressing a destabilized Cre (dgCre) theoretically active only upon trimethoprim (TMP) administration. TMP timing and dose influenced RC targeting efficiency, which was highest within the first three postnatal weeks, but specificity was low with many other spinal neurons also targeted. In addition, dgCre showed TMP-independent activity resulting in spontaneous recombination events that accumulated with age. Combining Calb1-dgCre with Parvalbumin (Pvalb) or Engrailed1 (En1) Flpo alleles in dual conditional systems increased cellular and timing specificity. Under optimal conditions, Calb1-dgCre/Pvalb-Flpo mice targeted 90% of RCs and few dorsal horn neurons; Calb1-dgCre/En1-Flpo mice showed higher specificity, but only a maximum of 70% of RCs targeted. Both models targeted neurons throughout the brain. Restricted spinal expression was obtained by injecting intraspinally AAVs carrying dual conditional genes. These results describe the first models to genetically target RCs bypassing development. |
| first_indexed | 2024-12-19T07:07:24Z |
| format | Article |
| id | doaj.art-af3e9dd9cd27406bb1dd54bc378acfda |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-19T07:07:24Z |
| publishDate | 2021-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-af3e9dd9cd27406bb1dd54bc378acfda2022-12-21T20:31:14ZengNature PortfolioScientific Reports2045-23222021-10-0111111710.1038/s41598-021-99333-6Genetic targeting of adult Renshaw cells using a Calbindin 1 destabilized Cre allele for intersection with Parvalbumin or Engrailed1Alicia R. Lane0Indeara C. Cogdell1Thomas M. Jessell2Jay B. Bikoff3Francisco J. Alvarez4Department of Physiology, Emory UniversityDepartment of Physiology, Emory UniversityDepartment of Biochemistry and Molecular Biophysics, Columbia UniversityDepartment of Developmental Neurobiology, St. Jude Children’s Research HospitalDepartment of Physiology, Emory UniversityAbstract Renshaw cells (RCs) are one of the most studied spinal interneurons; however, their roles in motor control remain enigmatic in part due to the lack of experimental models to interfere with RC function, specifically in adults. To overcome this limitation, we leveraged the distinct temporal regulation of Calbindin (Calb1) expression in RCs to create genetic models for timed RC manipulation. We used a Calb1 allele expressing a destabilized Cre (dgCre) theoretically active only upon trimethoprim (TMP) administration. TMP timing and dose influenced RC targeting efficiency, which was highest within the first three postnatal weeks, but specificity was low with many other spinal neurons also targeted. In addition, dgCre showed TMP-independent activity resulting in spontaneous recombination events that accumulated with age. Combining Calb1-dgCre with Parvalbumin (Pvalb) or Engrailed1 (En1) Flpo alleles in dual conditional systems increased cellular and timing specificity. Under optimal conditions, Calb1-dgCre/Pvalb-Flpo mice targeted 90% of RCs and few dorsal horn neurons; Calb1-dgCre/En1-Flpo mice showed higher specificity, but only a maximum of 70% of RCs targeted. Both models targeted neurons throughout the brain. Restricted spinal expression was obtained by injecting intraspinally AAVs carrying dual conditional genes. These results describe the first models to genetically target RCs bypassing development.https://doi.org/10.1038/s41598-021-99333-6 |
| spellingShingle | Alicia R. Lane Indeara C. Cogdell Thomas M. Jessell Jay B. Bikoff Francisco J. Alvarez Genetic targeting of adult Renshaw cells using a Calbindin 1 destabilized Cre allele for intersection with Parvalbumin or Engrailed1 Scientific Reports |
| title | Genetic targeting of adult Renshaw cells using a Calbindin 1 destabilized Cre allele for intersection with Parvalbumin or Engrailed1 |
| title_full | Genetic targeting of adult Renshaw cells using a Calbindin 1 destabilized Cre allele for intersection with Parvalbumin or Engrailed1 |
| title_fullStr | Genetic targeting of adult Renshaw cells using a Calbindin 1 destabilized Cre allele for intersection with Parvalbumin or Engrailed1 |
| title_full_unstemmed | Genetic targeting of adult Renshaw cells using a Calbindin 1 destabilized Cre allele for intersection with Parvalbumin or Engrailed1 |
| title_short | Genetic targeting of adult Renshaw cells using a Calbindin 1 destabilized Cre allele for intersection with Parvalbumin or Engrailed1 |
| title_sort | genetic targeting of adult renshaw cells using a calbindin 1 destabilized cre allele for intersection with parvalbumin or engrailed1 |
| url | https://doi.org/10.1038/s41598-021-99333-6 |
| work_keys_str_mv | AT aliciarlane genetictargetingofadultrenshawcellsusingacalbindin1destabilizedcrealleleforintersectionwithparvalbuminorengrailed1 AT indearaccogdell genetictargetingofadultrenshawcellsusingacalbindin1destabilizedcrealleleforintersectionwithparvalbuminorengrailed1 AT thomasmjessell genetictargetingofadultrenshawcellsusingacalbindin1destabilizedcrealleleforintersectionwithparvalbuminorengrailed1 AT jaybbikoff genetictargetingofadultrenshawcellsusingacalbindin1destabilizedcrealleleforintersectionwithparvalbuminorengrailed1 AT franciscojalvarez genetictargetingofadultrenshawcellsusingacalbindin1destabilizedcrealleleforintersectionwithparvalbuminorengrailed1 |