Papers by Paloma García-Bellido
Cellular and Molecular Biology, Nov 30, 2022
BMC Medical Genetics, May 2, 2019
Background: Mutations in the coding region of FOXP2 are known to cause speech and language impair... more Background: Mutations in the coding region of FOXP2 are known to cause speech and language impairment. However, it is not clear how dysregulation of the gene contributes to language deficit. Interestingly, microdeletions of the region downstream the gene have been associated with cognitive deficits. Methods: Here, we investigate changes in FOXP2 expression in the SK-N-MC neuroblastoma human cell line after deletion by CRISPR-Cas9 of two enhancers located downstream of the gene. Results: Deletion of any of these two functional enhancers downregulates FOXP2, but also upregulates the closest 3′ gene MDFIC. Because this effect is not statistically significant in a HEK 293 cell line, derived from the human kidney, both enhancers might confer a tissue specific regulation to both genes. We have also found that the deletion of any of these enhancers downregulates six well-known FOXP2 target genes in the SK-N-MC cell line. Conclusions: We expect these findings contribute to a deeper understanding of how FOXP2 and MDFIC are regulated to pace neuronal development supporting cognition, speech and language.
There is increasing evidence that mutations in the transcription factor FOXP2 impairs sensorimoto... more There is increasing evidence that mutations in the transcription factor FOXP2 impairs sensorimotor responses at the brain level. How some gene interactions produce afferentefferent circuits involving gabaergic and glutamergic populations of cells in different parts of the CNS is unclear. It is known that FOXP2 is expressed in a sensorimotor dopaminergic circuit, comprising the striatum, thalamus, deep cerebral cortical layers, the inferior olive and Purkinje cells of the cerebellum. Here we focus on a case of a subject A with speech and language disorders who has a chromosomal translocation t[7;11] affecting 7q31, the locus of FOXP2. Since there is substantial evidence that the neural basis for interval timing of fast movement changes, crucial for speech and language, may be regulated by these sensorimotor dopaminergic circuits, we focus here on how interval timing in the ms range is reproduced by A compared to the tutor, and a control C matched for sex, age, languages and education. It is found that A reproduces non-word sequences with significantly fewer dynamic changes than C. We discuss these findings relating them to the debatable hypothesis that the cerebellum may be more involved in the perception and production of sub-second intervals.
How exactly auditory stimulation initiates language comprehension is still unknown. Some lines of... more How exactly auditory stimulation initiates language comprehension is still unknown. Some lines of research point to the possible involvement of sub-cortical and cortical circuits, particularly through a functional network comprising the striatum-thalamus-cortex and cerebellum, in which FOXP2 is expressed. This network is thought to support the production and perception of time intervals. In order to evaluate if deficient perception and timing of short time interval words (SIW) in the range of 20 to 200 ms-crucial for timing correctly the integration of long time interval words (LIW) above 200ms-can increase the degree of difficulty in comprehension, we study here a 11 year old subject, A, with a break at some point in the 7q31 region, where FOXP2 is located. Using an audiovisual test with linguistically relevant SIWs contained in basic grammatical constructions and comparing A's difficulty score with that of a 4-11 year old normal population (NP) and that of a control matched in age, sex, schooling, language and socioeconomic background, it was found that A scored higher than a 6 year old but lower than a 7 year old. Subject A's anomalous degree of difficulty for specific constructions with SIWs is not consistent with these ages' performance, but rather suggests an anomalous impairment in perceiving and timing SIWs without which timing correctly the integration of LIWs might be disrupted.
The properties of Voiced Palatal Simplification (VPS) are of interest to any theory which investi... more The properties of Voiced Palatal Simplification (VPS) are of interest to any theory which investigates how linguistic elements match and what effects their matching triggers. This analysis supports models in cognitive linguistics where the selection for production is derived from the effects of an inhibitory process which lowers activation levels in one of the two matched linguistic items (Green, 1998; I'oulisse, 1997). It is argued on empirical grounds that VPS fails to be analysed as the movement of a Voiced Palatal (VP) in Phonetic Form (PF) to a site S where its voiced palatality is checked against that (>1 another VP in S, and its PF is subsequently deleted. Instead it is argued that the matching of VPs is a sufficient but not a necessary condition or VI'ti. Soune specific matchings do not flag their matching. VP matching is a local syntactic operation in the minimalist sense (Chomsky, 1995) Suhicct to operating in a c-command domain. VP matching cannot be activated inside an adjoined structure. It is suggested that all VP matchings fire activation of the matched sites in the form of a transcription. This transcription reaps all the Non-Phonetic Form (NPF) features of a matched VP onto a Phonetic Form (PF) which will be available for production. The activation levels of transcription are lowered or dampened by an inhibitory process in all sites where a successful matching has been initiated and flagged. These findings are relevant for theories of language development. SYNTAXIS 3 (2000) 75-109 www2.uhu.es/syntaxis
Estudios de Lingüística del Español (ELiEs), 2005
It is widely accepted in the scientific community that Language is produced by an organism, which... more It is widely accepted in the scientific community that Language is produced by an organism, which via its motor cortex, sets in motion vocal and hand-movements. Recent FMR studies show motor cortex activity for verbal motor tasks (Brown & Hagoort 2003:220). However, little is known about the specific biochemical mechanisms the brain activates or inhibits when using language. One of the most important discoveries of the XX century has been to realise that only approximately 30,000 genes can generate 20 billion neurons in a human brain and produce remarkably varied behaviour (Marcus 2004). Despite this neuronal explosion, it is revealed from imaging studies that only a small fraction of these neurons are activated for language production or interpretation and that regardless of the apparent complexity that any language may show, a human brain can effortlessly master it with little exposure to the environment. This fact may perhaps indicate that few genes and little environmental exposure may be necessary and sufficient to produce much variation in behaviour. What seems universally accepted by followers of neutral monism, is that knowing how gene regulation affects the physiology and biochemistry of brain cells, will eventually elucidate how linguistic effects emerge from these anatomical and biochemical interactions (Edelman & Tononi 2001, Marcus 2004)
Molecular Cytogenetics, Jun 10, 2015
Background: We report on a young female, who presents with a severe speech and language disorder ... more Background: We report on a young female, who presents with a severe speech and language disorder and a balanced de novo complex chromosomal rearrangement, likely to have resulted from a chromosome 7 pericentromeric inversion, followed by a chromosome 7 and 11 translocation. Results: Using molecular cytogenetics, we mapped the four breakpoints to 7p21.1-15.3 (chromosome position:
How exactly auditory stimulation initiates language comprehension is still unknown. Some lines of... more How exactly auditory stimulation initiates language comprehension is still unknown. Some lines of research point to the possible involvement of sub-cortical and cortical circuits, particularly through a functional network comprising the striatum-thalamus-cortex and cerebellum, in which FOXP2 is expressed. This network is thought to support the production and perception of time intervals. In order to evaluate if deficient perception and timing of short time interval words (SIW) in the range of 20 to 200 ms crucial for timing correctly the integration of long time interval words (LIW) above 200ms can increase the degree of difficulty in comprehension, we study here a 11 year old subject, A, with a break at some point in the 7q31 region, where FOXP2 is located. Using an audio-visual test with linguistically relevant SIWs contained in basic grammatical constructions and comparing A’s difficulty score with that of a 4-11 year old normal population (NP) and that of a control matched in ag...
ABSTRACTMutations in the coding region of FOXP2 are known to cause speech and language impairment... more ABSTRACTMutations in the coding region of FOXP2 are known to cause speech and language impairment. Microdeletions involving the region downstream the gene have been also associated to speech and cognitive deficits. We recently described a girl harbouring a complex chromosomal rearrangement with one breakpoint downstream the gene that might affect her speech and cognitive abilities via physical separation of distant regulatory DNA elements. In this study, we have used highly efficient targeted chromosomal deletions induced by the CRISPR/Cas9 genome editing tool to demonstrate the functionality of two enhancers, FOXP2-Eproximal and FOXP2-Edistal, located in the intergenic region between FOXP2 and its adjacent MDFIC gene. Deletion of any of these two functional enhancers in the neuroblastomic cell line SK-N-MC downregulates FOXP2 and decreases FOXP2 protein levels, conversely it upregulates MDFIC and increases MDFIC protein levels. This suggests that both regulatory elements may be sha...
How exactly auditory stimulation initiates language comprehension is still unknown. Some lines of... more How exactly auditory stimulation initiates language comprehension is still unknown. Some lines of research point to the possible involvement of sub-cortical and cortical circuits, particularly through a functional network comprising the striatum-thalamus-cortex and cerebellum, in which FOXP2 is expressed. This network is thought to support the production and perception of time intervals. In order to evaluate if deficient perception and timing of short time interval words (SIW) in the range of 20 to 200 ms - crucial for timing correctly the integration of long time interval words (LIW) above 200ms - can increase the degree of difficulty in comprehension, we study here a 11 year old subject, A, with a break at some point in the 7q31 region, where FOXP2 is located. Using an audio-visual test with linguistically relevant SIWs contained in basic grammatical constructions and comparing A’s difficulty score with that of a 4-11 year old normal population (NP) and that of a control matched i...
Actas del simposio internacional El legado de Rafael Lapesa, 2008, ISBN 978-84-482-5103-1, págs. 159-163, 2008
Este trabajo fue leído en el XIII Simposio de la Sociedad Española de Lingüística celebrado en Ba... more Este trabajo fue leído en el XIII Simposio de la Sociedad Española de Lingüística celebrado en Barcelona en diciembre de 1983 con el título de «La ordenación de las reglas y el ciclo fonológico».In this paper it has been shown that certain irregularities in the verbal system of Spanish do not need to be explained as a consequence of stipulating two opposite orderings in the Spanish Grammar. We have offered a new alternative within a much more restricted Phonological framework. This alternative makes crucial use of Universal Principles such as The Elsewhere Condition and Level-ordered Morphology. The goal of this paper is to show that only one ordering seems to be possible, moreover that none of the criteria used so far to justify «double ordering» such as «paradigmatic uniformity» and «paradigmatic irregularity» or «preferable» orderings are required any longer in the Theory. This lexical approach has led us to give account not only of irregularities in the verbal Spanish system but...
Syntaxis, 2000
The properties of Voiced Palatal Simplification (VPS) are of interest to any theory which investi... more The properties of Voiced Palatal Simplification (VPS) are of interest to any theory which investigates how linguistic elements match and what effects their matching triggers. This analysis supports models in cognitive linguistics where the selection for production is derived from the effects of an inhibitory process which lowers activation levels in one of the two matched linguistic items (Green, 1998; I'oulisse, 1997). It is argued on empirical grounds that VPS fails to be analysed as the movement of a Voiced Palatal (VP) in Phonetic Form (PF) to a site S where its voiced palatality is checked against that (>1 another VP in S, and its PF is subsequently deleted. Instead it is argued that the matching of VPs is a sufficient but not a necessary condition or VI'ti. Soune specific matchings do not flag their matching. VP matching is a local syntactic operation in the minimalist sense (Chomsky, 1995) Suhicct to operating in a c-command domain. VP matching cannot be activated inside an adjoined structure. It is suggested that all VP matchings fire activation of the matched sites in the form of a transcription. This transcription reaps all the Non-Phonetic Form (NPF) features of a matched VP onto a Phonetic Form (PF) which will be available for production. The activation levels of transcription are lowered or dampened by an inhibitory process in all sites where a successful matching has been initiated and flagged. These findings are relevant for theories of language development. SYNTAXIS 3 (2000) 75-109 www2.uhu.es/syntaxis
Table S1. Oligonucleotide and sgRNA sequences used in this study. (DOCX 15 kb)
Figure S6. Detailed view of an ENCODE UCSC genome-browser snapshot showing bar graphs with a deta... more Figure S6. Detailed view of an ENCODE UCSC genome-browser snapshot showing bar graphs with a detailed representation of the locations of FOXP2 and MDFIC genes, H3K27Ac and DNA clusters in human cell lines. The squared regions in black show the locations of FOXP2-Eproximal and FOXP2-Edistal. The red squared tracks show the alignment result between humans and bats. (JPG 761 kb)
Figure S5. RT-qPCR analysis of six HEK293 cell clones with FOXP2-Eproximal or FOXP2-Edistal delet... more Figure S5. RT-qPCR analysis of six HEK293 cell clones with FOXP2-Eproximal or FOXP2-Edistal deletions. Samples are normalized to the average FOXP2 (left) or MDFIC (right) signal between three HEK293 replicates transfected with the pLV-U6#xH1#y-C9G empty vector. Levels of expression of FOXP2 and MDFIC are represented by the fold change relative to that of empty vector control cell line, which were normalized to 1. WT/WT: wild type, Δ/Δ: homozygous deletion, WT/Δ: heterozygous deletion. (JPG 373 kb)
Figure S4. PCR analysis. Two oligos flanking the deleted regions were used to amplify the genomic... more Figure S4. PCR analysis. Two oligos flanking the deleted regions were used to amplify the genomic DNA from several mutant representative HEK293 and SK-N-MC clones. Black triangles show the size of the PCR products. Black or white asterisks show respectively the clones harbouring a homozygous or heterozygous deletion included in this study. M: molecular weight marker, WT/WT: wild type, Δ/Δ: homozygous deletion, WT/Δ: heterozygous deletion. (JPG 1181 kb)
Figure S3. Indel spectrum determined by TIDE of the off-target sites compared with indel frequenc... more Figure S3. Indel spectrum determined by TIDE of the off-target sites compared with indel frequencies of the control sample. Each module represents the TIDE analysis of one sgRNA in a bulk cell population electroporated with each of the single-guide-Cas9 encoded plasmids. Each bar graph represents an indel event with an estimation of the percentage of the population exhibiting this particular event. On-target and potential off-target sequences are represented on top of each module and mismatched bases are shown in red. Light-red bars represent the wild type situation, bright-red bars represent significant indel events and black bars represent non-significant differences. P-values according to Pearson's chi-squared test. Decomposition was limited to indels of size 0–10, hence larger indels could not be detected. R2 represent a quality measurement of the sequence reads. Indel % is represented at the top left site each module. (JPG 1041 kb)
Figure S2. Indel spectrum determined by TIDE of the on-target sites compared with indel frequenci... more Figure S2. Indel spectrum determined by TIDE of the on-target sites compared with indel frequencies of the control sample. Each module represents the TIDE analysis of one sgRNA in a bulk cell population electroporated with each of the single-guide-Cas9 encoded plasmids. Each bar graph represents an indel event with an estimation of the percentage of the population exhibiting this particular event. Light-red bars represent the wild type control DNA sequence, bright-red bars represent significant indel events and black bars represent non-significant differences. P-values according to Pearson's chi-squared test. Decomposition was limited to indels of size 0–10, hence larger indels could not be detected. R2 represent a quality measurement of the sequence reads. Indel % is represented at the top left site each module. (JPG 1103 kb)
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Papers by Paloma García-Bellido